|
| Status |
Public on Aug 05, 2013 |
| Title |
HL60-undifferentiated |
| Sample type |
SAGE |
| Anchor |
NlaIII
|
| Tag Count |
33636 |
| Tag Length |
17 |
| |
|
| Source name |
Acute promyelocytic lukemia HL-60 cell line (Catalog No.CCL-240)
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HL60
cell type: Acute promyelocytic lukemia
status: Undifferentiated
|
| Treatment protocol |
HL60 cells were differentiated with 1.3% DMSO (Sigma catalog No. D4540) in complete media for 6-8 days with media change every 3rd day. KG60 cells were differentiated with PMA (100ng/ml) for 5 days.
|
| Growth protocol |
HL60 and KG1 cell were maintained in complete Iscoves Modified Medium (ATCC catalog No.30-2005) supplemented with 10% fetal bovine serum (ATCC catalog No. 30-2020), penicillin (100U/ml), and streptomycin (100µg/ml), at 37 degrees C and 5% CO2. Neutrophiles were maintained in Krebs-HEPES + BSA.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total cellular RNA was isolated from differentiated and from undifferentiated cell lines and human naïve neutrophils using tri-Reagent (MRC, Cincinati, OH) following manufacturer’s instructions. RNA integrity was routinely checked using 500 ng/ml of RNA on 1% denaturing agarose gel. SAGE was performed according to the Serial Analysis of Gene Expression detailed Protocol Version C, with Nla III as the digestion enzyme, Mme as the tagging enzyme.
|
| |
|
| Data processing |
Transformants were analyzed by colony PCR. Approximately 4000-5000 clones for each library were cycle sequenced using M13 forward primer and analyzed on Applied Biosystems DNA Sequencer. Approximately 20-25 individual tags were produced per clone. Sequences were analyzed using SAGE analysis software version 4.5 (Johns Hopkins University, Baltimore, MD, USA) and annotated using the downloadable reference sequence database SAGEmap.
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| |
|
| Submission date |
Dec 31, 2012 |
| Last update date |
Aug 05, 2013 |
| Contact name |
Anason Halees |
| E-mail(s) |
ahalees@kfshrc.edu.sa
|
| Organization name |
King Faisal Specialist Hospital & Research Centre
|
| Department |
Research Center
|
| Lab |
Cell Biology
|
| Street address |
P.O.Box 3354
|
| City |
Riaydh |
| ZIP/Postal code |
11211 |
| Country |
Saudi Arabia |
| |
|
| Platform ID |
GPL1485 |
| Series (1) |
| GSE43211 |
Differentiated vs. undifferentiated promyelocytic cell lines (HL-60 and KG-1) and Neutrophiles |
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